Optical assessment of motoneuron function in a “twenty-four-hour” acute spinal cord slice model from fetal rats

Metzger, Friedrich; Klapproth, Nadine; Kulik, Anna; Sendtner, Michael; Ballanyi, Klaus

doi:10.1016/j.jneumeth.2004.07.016

Cited by 9 publications

(10 citation statements)

References 75 publications

(137 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Rhodamine-123, which is trapped inside mitochondria at autoquenching concentrations (A), is released into the cytosol under ischemic conditions described, resulting in a strong increase of the fluorescence signal (B). The Rhodamine-123 signal (thought to be associated with MPTP opening 24 is consistent with dose-response-relationships, kinetics and amplitudes of calcium transients shown in Figure 2 vivo effect on the formation of β-amyloid plaques in a genderspecific fashion in an aggressive model of cerebral amyloidosis. 19,20 The calcium-dependent formation of the MPTP is considered to underlie the initiation of the intrinsic apoptotic pathway in a variety of neurodegenerative conditions, 26,27 in particular with regard to Alzheimer's disease.…”

Section: Discussionsupporting

confidence: 78%

“…sequestered inside mitochondria at autoquenching concentrations. Upon activation of the MPTP by ischemia, the dye is released into the cytosol generating a strong fluorescent signal, 24 as shown in Figure 4A and B for a representative neuron from the ESC model. We observe a dose-dependent inhibition of this type of signal by Dutasteride (and related compounds, not shown).…”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

A Connection between the Mitochondrial Permeability Transition Pore, Autophagy, and Cerebral Amyloidogenesis

et al. 2008

View full text Add to dashboard Cite

In a drug reprofiling attempt, we explored novel neuroprotective properties of 4-azasteroids by synthesizing chemical affinity tags capturing adenine nucleotide translocator-1, as a potential target. Dutasteride inhibits the mitochondrial transition pore and induces an increase of autophagosomal structures in human cell lines. In vivo, a surprising reduction of the beta-amyloid plaque load in a model for cerebral amyloidosis appears to connect release of neurotoxic peptides, mitochondrial apoptosis and autophagy.

show abstract

Section: Discussionsupporting

confidence: 78%

Section: Resultsmentioning

confidence: 99%

A Connection between the Mitochondrial Permeability Transition Pore, Autophagy, and Cerebral Amyloidogenesis

et al. 2008

View full text Add to dashboard Cite

show abstract

“…Because of the large electrochemical gradient across the mitochondrial inner membrane, R123 accumulates preferentially in the matrix of this organelle (12, 38). This property of R123 has been used to quantify the mitochondrial potential (Δψ) in living cells as well as isolated mitochondria by means of fluorescence microscopy, fluorimetry, flow cytometry, and confocal microscopy (1, 37, 38, 51–53). Here, we used the binary image as a mask for spatial quantification of the R123 fluorescence intensity in the “RAW” image.…”

Section: Discussionmentioning

confidence: 99%

Simultaneous quantitative measurement and automated analysis of mitochondrial morphology, mass, potential, and motility in living human skin fibroblasts

et al. 2005

View full text Add to dashboard Cite

Background: Understanding the interdependence of mitochondrial and cellular functioning in health and disease requires detailed knowledge about the coupling between mitochondrial structure, motility, and function. Currently, no rapid approach is available for simultaneous quantification of these parameters in single living cells. Methods: Human skin fibroblasts were pulse-loaded with the mitochondria-selective fluorescent cation rhodamine 123. Next, mitochondria were visualized using video-rate (30 Hz) confocal microscopy and real-time image averaging. To highlight the mitochondria, the acquired images were binarized using a novel image processing strategy. Results: Our approach enabled rapid and simultaneous quantification of mitochondrial morphology, mass, potential, and motility. It was found that acute inhibition of

show abstract

“…damin-123 (Metzger et al, 2005) and in Figure 4G, a typical Fura-2 calcium image of neurons from hESC after exposure to 10 11M glutamate is shown as a measure of functional viability. (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Slices were loaded with rhodamine-123 were excited at 480 nm and emission was monitored at 565 nm. Mitochondrial rhodamine-123 accumulation and quenching results in a fiuorescence decrease, whereas depolarisation provokes signal dequenching and an increase of fiuorescence after mitochondrial dye release (Metzger et al, 2005).…”

Section: Assessment 01 In Vitro Functionality 01 Hes-derived Neurons mentioning

confidence: 99%

Neuronal cell culture from human embryonic stem cells as in vitro model for neuroprotection

Schrattenholz

Klemm²

2007

ALTEX

View full text Add to dashboard Cite

In the context of efficacy testing ofpharmacological compounds in animal models, replacement of some of these models with a relevant human in vitro system appears attractive, in particular with regard to large scale screening. Here, we show resultsfrom initial phases of a project, which attempts to explore the outstanding potential of human embryonie stem cell (hESC)based in vitro models with special regard to neuronal stress as a potential replacement of animal models for human neurodegenerative diseases. We show the functionality of neurons derived from hESC precursors by calcium imaging, mitochondrial potential measurements and Western blots and moreover demonstrate that this model reproduces crucial mechanistic aspects observed during ischemia and excitotoxicity that are thought to be at the core of some neurodegenerative diseases. Also, the broader possibilities for refining surrogate molecular information emerging from the detailed analysis of this model are discussed. Zusammenfassung: Stammzellbasierte menschliche in vitro Modelle zum Ersatz der Wirksamkeitsprüfung bei Tieren Im Kontext von Wirksamkeitsstudien von pharmakologischen Formulierungen und Medikamentenkandidaten in Tiermodellen besteht das starke Bedürfnis Ersatzmodelle auf der Basis menschlicher in vitro Systeme zu entwickeln. Wir zeigen hier Ergebnisse aus den frühen Phasen eines Projekts, welches das besondere Potential humaner embryonaler Stammzell-(hESC)-Modelle als entsprechende in vitro Systeme erforschen möchte, mit einem speziellem Fokus auf neuronalen Stress und mit dem Ziel Tiermodelle für humane neurodegenerative Krankheiten zu ersetzen. Wir konnten die Funktionalität von Neuronen, die aus hESC Vorläufern differenziert wurden, durch Calcium Imaging, Potentialmessungen der mitochondrialen Membran und Western Blots zeigen. Wir konnten auch nachweisen, dass das Modell wesentliche mechanistische Aspekte, wie sie während der Ischämie oder unter erregungstoxischen Bedingungen auftreten, gut widerspiegelt. Es wird vermutet, dass diese Prozesse einer Reihe von menschlichen neurodegenerativen Krankheiten zugrunde liegen. Auch weitergehende Möglichkeiten zur Entwicklung besserer und genauerer; auch molekularer Endpunkte, die sich aus der detaillierten Analyse dieser Modelle ergeben könnten, werden diskutiert.

show abstract

Optical assessment of motoneuron function in a “twenty-four-hour” acute spinal cord slice model from fetal rats

Cited by 9 publications

References 75 publications

A Connection between the Mitochondrial Permeability Transition Pore, Autophagy, and Cerebral Amyloidogenesis

A Connection between the Mitochondrial Permeability Transition Pore, Autophagy, and Cerebral Amyloidogenesis

Simultaneous quantitative measurement and automated analysis of mitochondrial morphology, mass, potential, and motility in living human skin fibroblasts

Neuronal cell culture from human embryonic stem cells as in vitro model for neuroprotection

Contact Info

Product

Resources

About